scholarly journals DETECTION AND MOLECULAR CHARACTERIZATION OF ODONTOGLOSSUM RINGSPOT VIRUS (ORSV) JAVA AND BALI ISOLATES

2015 ◽  
Vol 2 (1) ◽  
pp. 112
Author(s):  
Mahfut , ◽  
Budi Setiadi Daryono ◽  
Trijoko ,
Keyword(s):  
Coat Protein ◽  
Ornamental Plants ◽  
Ringspot Virus ◽  
Limiting Factor ◽  
Pathogenicity Test ◽  
Virus Infections ◽  
Rt Pcr ◽  
Chenopodium Amaranticolor ◽  
Odontoglossum Ringspot Virus ◽  
Cp Gene

<p>Orchids are one of the most important ornamental plants and cultivated in tropical countries, including in Indonesia. Virus infections become an important limiting factor in orchids cultivation because it causes significant losses of the plants. Odontoglossum ringspot virus (ORSV) is one of the most reported viruses infecting orchids and spread widely in the world. The common symptoms are mosaic with line pattern and necrotic ringspot on leaf surface, and also color breaking on flowers. The purpose of this research was to find out the occurrence of ORSV infecting orchids in Java and Bali, genetic relationship among ORSV isolates based on similarities of coat protein (CP) gene, and to analyze the pathogenicity test of ORSV Java and Bali isolates. Survey and samples collection were conducted in seven locations cultivating orchids in Java and Bali. Detection with Reverse Transcription-Polymerase Chain Reaction (RT-PCR) showed that ORSV was infected three leaf samples of Phalaenopsis sp., called ORSV BOC, ORSV KRB and ORSV TNBB isolates. The results showed 474 bp-amplified DNA band as the expression of ORSV CP gene. Phylogenetic analysis based on nucleotide sequences of CP gene showed that ORSV BOC have similarity close to ORSV Germany, whereas ORSV KRB and ORSV TNBB leads to speciation that possible to be a new strain. Pathogenicity test using various healthy plants showed that ORSV BOC may infected and cause systemic symptoms on Chenopodium amaranticolor, Nicotiana tabacum, Dendrobium sp., Cymbidium sp., Chattleya sp., Phalaenopsis sp., Liparis sp., Spatthoglotis sp. and Pectelis sussanae (L.) Raf.</p><p><br /><strong>Keywords:</strong> ORSV, orchids, coat protein, RT-PCR</p>

scholarly journals SURVEI Odontoglossum ringspot virus (ORSV) YANG MENGINFEKSI ANGGREK ALAM TROPIS DI INDONESIA (SURVEY ON THE OCCURRENCE OF Odontoglossum ringspot virus (ORSV) INFECTING NATURE TROPICAL ORCHIDS IN INDONESIA)

2017 ◽  
Vol 20 (1) ◽  
pp. 1
Author(s):  
Mahfut Mahfut ◽  
Budi Setiadi Daryono ◽  
Tri Joko ◽  
Susamto Somowiyarjo
Keyword(s):  
Viral Infections ◽  
Serological Test ◽  
Ornamental Plants ◽  
Ringspot Virus ◽  
Limiting Factor ◽  
Natural Forests ◽  
Virus Infections ◽  
Odontoglossum Ringspot Virus ◽  
Total Incidence ◽  
Das Elisa

ABSTRACTNatural orchids are one of the important ornamental plants that were cultivated in tropical countries, including Indonesia. Virus infections has been important limiting factor in orchids cultivation because it decreases the orchids quality. Odontoglossum ringspot virus (ORSV) is one of the most reported virus infecting orchids which spread widely in the world. During 2010–2014 surveys of viral infections were conducted in Indonesia. The orchids were found infected by virus, showed symptoms of mosaic, mottle, chlorotic, necrotic, streak, wilting leaf, and ringspot on leaf surface. Detection with serological test DAS-ELISA showed only 11 from 125 samples were infected by ORSV with total incidence of 8,8%. Nine leaf samples of Phalaenopsis sp. were infected, respectively. Thus, it was concluded that Phalaenopsis is orchids genus which is the most abundantly and susceptibly infected by ORSV. The results proved that ORSV have entered and spread widely by infected orchids in orchids landscape (nursery), semi-natural forests (botanical gardens), and natural forest (national park) throughout Indonesia. This is the first report of ORSV infecting natural tropical orchids in Indonesia.Keywords: DAS-ELISA, Indonesia, nature tropical orchids, ORSVINTISARIAnggrek alam merupakan salah satu kekayaan flora asli negara tropis, termasuk Indonesia. Keberadaan di habitat aslinya sudah sangat berkurang yang disebabkan kerusakan hutan dan adanya penyakit. Infeksi virus masih menjadi faktor pembatas terpenting dalam budidaya dan pengembangan potensi anggrek alam. Odontoglossum ringspot virus (ORSV) adalah salah satu jenis virus yang dilaporkan paling banyak menginfeksi anggrek serta memiliki penyebaran yang luas di dunia. Selama 2010-2014 telah dilakukan survei lapangan terhadap infeksi virus di Indonesia. Beberapa anggrek yang ditemukan terinfeksi oleh virus menunjukkan gejala berupa mosaik, belang, klorosis, nekrosis, streak, daun layu, dan bercak cincin pada permukaan daun. Deteksi dengan uji serologis DAS-ELISA menunjukkan bahwa 11 dari 125 sampel terinfeksi oleh ORSV dengan total kejadian 8,8%. Masing-masing sembilan dari total sampel daun terinfeksi merupakan Phalaenopsis sp. Hal ini menjelaskan bahwa Phalaenopsis adalah genus anggrek yang paling cocok dan rentan terhadap infeksi ORSV. Hasil penelitian survei kejadian infeksi Odontoglossum ringspot virus (ORSV) membuktikan bahwa virus ini telah masuk dan menyebar secara luas oleh anggrek-anggrek alam tropis yang terinfeksi di pertamanan anggrek (nurseri), hutan semi-alami (kebun raya), dan hutan alam (taman nasional) di seluruh wilayah Indonesia. Ini merupakan laporan pertama mengenai infeksi ORSV terhadap anggrek-anggrek alam tropis di Indonesia.Kata kunci: anggrek alam tropis, DAS-ELISA, Indonesia, ORSV

scholarly journals Caracterização de uma nova estirpe do Tomato mosaic virus isolada de tomateiro no Estado de São Paulo

2003 ◽  
Vol 28 (6) ◽  
pp. 602-607 ◽  
Author(s):  
Silvia R. Moreira ◽  
Marcelo Eiras ◽  
Alexandre L.R. Chaves ◽  
Silvia R. Galleti ◽  
Addolorata Colariccio
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Nicotiana Sylvestris ◽  
Sao Paulo ◽  
Ringspot Virus ◽  
Tomato Mosaic Virus ◽  
Rt Pcr ◽  
Chenopodium Amaranticolor ◽  
Odontoglossum Ringspot Virus

Um vírus isolado em Guaratinguetá, SP, de tomateiro (Lycoporsicon esculentum) 'Santa Clara' com sintomas característicos de virose, foi estudado por meio de plantas indicadoras e de hospedeiras diferenciais pertencentes a linhagens homozigotas de tomateiro, ensaios de estabilidade in vitro, purificação, contrastação negativa, testes sorológicos de ELISA-PTA e imunomicroscopia eletrônica, utilizando-se anti-soros contra diferentes vírus do gênero Tobamovirus. O isolado infetou plantas de espécies de amarantáceas, quenopodiáceas e solanáceas. Plantas de Chenopodium amaranticolor reagiram com sintomas locais e sistêmicos; Nicotiana sylvestris e N. rustica reagiram com lesões locais e a linhagem homozigota de tomateiro Tm-2 mostrou-se imune ao vírus. Nas preparações purificadas de contrastação negativa, foram observadas partículas rígidas e alongadas com cerca de 300 nm. O isolado foi identificado como um tobamovírus, com anti-soros contra o Tomato mosaic virus (ToMV) e Tobacco mosaic virus (TMV). As hospedeiras diferenciais indicaram se tratar de ToMV. Por meio de RT-PCR, com oligonucleotídeos para o gene da capa protéica de espécies do gênero Tobamovirus do subgrupo 1, amplificaram-se fragmentos com 850 pb que foram clonados e seqüenciados. A similaridade de nucleotídeos e aminoácidos deduzidos variou entre 85 e 91% quando a seqüência do ToMV-SP foi comparada com outras sequências de ToMV, 75 e 83% quando comparada com as do TMV e 67 e 72% quando comparada com a do Odontoglossum ringspot virus (ORSV). As comparações com outras espécies de tobamovírus apresentaram valores de similaridade inferiores a 65%. Confirmou-se a identidade dos vírus como sendo uma nova estirpe do ToMV.

Prevalence and Molecular Characterization Coat Protein Gene of Tobacco streak virus Causing Peanut Stem Necrosis Disease in Coastal and Rayalaseema Districts of Andhra Pradesh, South-India

2021 ◽  
Author(s):  
K. Saratbabu ◽  
K. Vemana ◽  
A.K. Patibanda ◽  
B. Sreekanth ◽  
V. Srinivasa Rao
Keyword(s):  
Coat Protein ◽  
Molecular Characterization ◽  
Andhra Pradesh ◽  
Disease Incidence ◽  
Tobacco Streak Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Streak Virus ◽  
Rt Pcr ◽  
Cp Gene ◽  
Stem Necrosis

Background: Peanut stem necrosis disease (PSND) caused by Tobacco streak virus (TSV) is a major constraint for groundnut production in Andhra Pradesh (A.P.). However, studies on prevalence and spread of the disease confined to only few districts of A.P. with this background current study focused on incidence and spread of the disease in entire state of A.P. Further an isolate of TSV occurring in A.P. characterized on the basis of genetic features by comparing with other TSV isolates originated from different hosts and locations from world.Methods: Roving survey was conducted during kharif 2017-18 in groundnut growing districts of Andhra Pradesh (A.P.) for peanut stem necrosis disease incidence. Groundnut plants showing PSND symptoms were collected and tested with direct antigen coating enzyme linked immunosorbent assay (DAC-ELISA). Groundnut samples found positive by ELISA once again tested by reverse transcription polymerase chain reaction (RT-PCR). The representative TSV-GN-INDVP groundnut isolate from Prakasham district was maintained on cowpea seedlings by standard sap inoculation method in glasshouse for further molecular characterization. The Phylogenetic tree for coat protein (CP) gene was constructed using aligned sequences with 1000 bootstrap replicates following neighbor-joining phylogeny.Result: Thirty-eight (52.7%) of seventy-two groundnut samples collected from different locations in A.P were given positive reaction to TSV by DAC-ELISA. For the first time, PSND incidence observed in coastal districts (Krishna, Guntur, Sri Pottisriramulu Nellore, Prakasham) of A.P. Maximum PSND incidence recorded from Bathalapalli (22.2%) and the minimum incidence in Mulakalacheruvu (4.1%). The coat protein (CP) gene of TSV-GN-INDVP groundnut isolate was amplified by RT-PCR and it shared maximum per cent nucleotide identity (97.51-98.62%) with TSV isolates from groundnut and other different crops reported in India. All Indian isolates cluster together irrespective of crop and location based on the phylogenetic analysis.

scholarly journals First Report of Cymbidium mosaic virus and Odontoglossum ringspot virus in Orchids in Mexico

Plant Disease ◽  
2012 ◽  
Vol 96 (3) ◽  
pp. 464-464
Author(s):  
A. G. Soto-Valladares ◽  
R. De La Torre-Almaraz ◽  
B. Xoconostle-Cazares ◽  
R. Ruíz-Medrano
Keyword(s):  
Mosaic Virus ◽  
Pcr Amplification ◽  
Positive Sample ◽  
Ringspot Virus ◽  
Rdrp Gene ◽  
Mixed Infections ◽  
Cymbidium Mosaic Virus ◽  
First Report ◽  
Odontoglossum Ringspot Virus ◽  
Cp Gene

In 2010, a survey for viral diseases in commercial, orchid-producing greenhouses was carried out in Morelos, Mexico. Many symptomatic plants were observed. The most common leaf symptoms were yellow mottle, yellow streaks, and chlorotic and necrotic ringspots. Leaf samples were collected from eight symptomatic plants from the following genera: Encyclia, Oncidium, Shomburghia, Brassia, Guarianthe, Cattleya, Epidendrum, Vanilla, Xilobium, Laelia, and Brassocattleya. Samples were tested using double-antibody sandwich (DAS)-ELISA (Agdia, Elkhart, IN) with antiserum for Cymbidium mosaic virus (CymMV), Odontoglossum ringspot virus (ORSV), Cymbidium ringspot mosaic virus, and Tobacco mosaic virus (TMV) and a general antiserum for potyviruses. At least one plant from each genus was positive to CymMV and ORSV as individual or mixed infections. Encyclia and Laelia plants were the most frequently found with mixed infections by both viruses. All genera were negative for TMV and potyviruses. Total RNA extracts were obtained from all ELISA-positive samples by a modified silica capture protocol (2). Reverse transcription (RT)-PCR was carried out with general polymerase (RdRp) gene primers corresponding to the Potexvirus group (3) and specific primers for the coat protein gene (CP) of CymMV and ORSV (1). The PCR amplification from a positive sample of each genus was resolved in agarose gels. Amplification products of the expected size were obtained for CymMV and ORSV. Five CymMV RdRp gene clones from five different plants of Laelia (GenBank Accession Nos. HQ393958, HQ393959, HQ393960, HQ393961, and HQ393962), two CP gene clones of CP gene of CymMV from two different plants of Oncidium (GenBank Accession Nos. HQ393956 and HQ393957), and three CP clones of CP of ORSV from three different plants of Encyclia (GenBank Accession Nos. HQ393953, HQ393954, and HQ393955) were sequenced. The nucleotide sequences of the Mexican orchid CymMV isolates were 96 to 97% identical to CymMV sequences in the GenBank, while those of ORSV were 99 to 100% identical to deposited ORSV sequences. To our knowledge, this is the first report of CymMV and ORSV in orchids in Mexico, which are two of the most important quarantine virus in orchids in Mexico. References: (1) P. Ajjikuttira et al. J. Gen. Virol. 86:1543, 2005. (2) J. R. Thompson et al. J. Virol. Methods 111:85, 2003. (3) R. A. A. van der Vlugt and M. Berendsen. Eur. J. Plant Pathol. 108:367, 2002.

scholarly journals Engineered Resistance Against Papaya ringspot virus in Venezuelan Transgenic Papayas

Plant Disease ◽  
2004 ◽  
Vol 88 (5) ◽  
pp. 516-522 ◽  
Author(s):  
Gustavo Fermin ◽  
Valentina Inglessis ◽  
Cesar Garboza ◽  
Sairo Rangel ◽  
Manuel Dagert ◽  
...  
Keyword(s):  
Amino Acid ◽  
Agrobacterium Tumefaciens ◽  
Coat Protein ◽  
Amino Acid Sequence ◽  
Sequence Similarity ◽  
Ringspot Virus ◽  
Amino Acid Sequence Similarity ◽  
Papaya Ringspot Virus ◽  
Local Varieties ◽  
Cp Gene

Local varieties of papaya grown in the Andean foothills of Mérida, Venezuela, were transformed independently with the coat protein (CP) gene from two different geographical Papaya ringspot virus (PRSV) isolates, designated VE and LA, via Agrobacterium tumefaciens. The CP genes of both PRSV isolates show 92 and 96% nucleotide and amino acid sequence similarity, respectively. Four PRSV-resistant R0 plants were intercrossed or selfed, and the progenies were tested for resistance against the homologous isolates VE and LA, and the heterologous isolates HA (Hawaii) and TH (Thailand) in greenhouse conditions. Resistance was affected by sequence similarity between the transgenes and the challenge viruses: resistance values were higher for plants challenged with the homologous isolates (92 to 100% similarity) than with the Hawaiian (94% similarity) and, lastly, Thailand isolates (88 to 89% similarity). Our results show that PRSV CP gene effectively protects local varieties of papaya against homologous and heterologous isolates of PRSV.

scholarly journals Virus-like particles assemble in plants and bacteria expressing the coat protein gene of Indian peanut clump virus

Microbiology ◽  
2000 ◽  
Vol 81 (1) ◽  
pp. 267-272 ◽  
Author(s):  
C. Bragard ◽  
G. H. Duncan ◽  
S. V. Wesley ◽  
R. A. Naidu ◽  
M. A. Mayo
Keyword(s):  
Coat Protein ◽  
Gene Transcript ◽  
Bacterial Cells ◽  
Rt Pcr ◽  
Virus Like Particles ◽  
Virus Particles ◽  
Untranslated Sequence ◽  
Cp Gene ◽  
Virus Rna ◽  
Particle Length

cDNA copies of the coat protein (CP) gene of Indian peanut clump virus (IPCV)-H were introduced into cells of Nicotiana benthamiana or Escherichia coli by transformation with vectors based on pROKII or pET respectively. In both plant and bacterial cells, IPCV CP was expressed and assembled to form virus-like particles (VLP). In plant extracts, the smallest preponderant particle length was about 50 nm. Other abundant lengths were about 85 and about 120 nm. The commonest VLP length in bacterial extracts was about 30 nm. Many of the longer VLP appeared to comprise aggregates of shorter particles. The lengths of the supposed ‘monomer’ VLP corresponded approximately to those expected for encapsidated CP gene transcript RNA. Immunocapture RT–PCR, using primers designed to amplify the CP gene, confirmed that the VLP contained RNA encoding IPCV-H CP. The results show that encapsidation does not require the presence of the 5′-terminal untranslated sequence of the virus RNA and suggest that if there is an ‘origin of assembly’ motif or sequence, it lies within the CP gene. When transgenic plants expressing IPCV-H CP were inoculated with IPCV-L, a strain that is serologically distinct from IPCV-H, the virus particles that accumulated contained both types of CP.

scholarly journals Reciprocal function of movement proteins and complementation of long-distance movement of Cymbidium mosaic virus RNA by Odontoglossum ringspot virus coat protein

2005 ◽  
Vol 86 (5) ◽  
pp. 1543-1553 ◽  
Author(s):  
Prabha Ajjikuttira ◽  
Chiang-Shiong Loh ◽  
Sek-Man Wong
Keyword(s):  
Coat Protein ◽  
Transgenic Plants ◽  
Mosaic Virus ◽  
Cell Movement ◽  
Ringspot Virus ◽  
Deficient Mutant ◽  
Long Distance ◽  
Cymbidium Mosaic Virus ◽  
Odontoglossum Ringspot Virus ◽  
Long Distance Movement

Complementation of movement and coat proteins of the orchid-infecting potexvirus Cymbidium mosaic virus (CymMV) and tobamovirus Odontoglossum ringspot virus (ORSV) was investigated. Nicotiana benthamiana, which is susceptible to both CymMV and ORSV, was used as a model system. Four transgenic lines, each harbouring one of the movement protein (MP) or coat protein (CP) genes of CymMV or ORSV, were constructed. The MP of CymMV consists of three overlapping open reading frames, together called the triple-gene block (TGB). CymMV and ORSV mutants, each carrying an inactivated MP or CP, were generated from the respective biologically active full-length cDNA clones. Complementation was studied by infecting transgenic plants with in vitro transcripts generated from these mutants. The cell-to-cell movement of a movement-deficient CymMV was restored in transgenic plants carrying the ORSV MP transgene. Similarly, CymMV TGB1 transgenic plants were able to rescue the cell-to-cell movement of a movement-deficient ORSV mutant. ORSV CP transgenic plants supported systemic movement of a CymMV CP-deficient mutant. However, in these plants, neither encapsidation of CymMV RNA with ORSV CP nor CymMV CP expression was detected. Long-distance movement of an ORSV CP-deficient mutant was not supported by CymMV CP. The complementation of MPs and CPs of CymMV and ORSV facilitates movement of these viruses in plants, except for long-distance movement of ORSV RNA by CymMV CP.

Genetic variability in the coat protein genes of two orchid viruses: Cymbidium mosaic virus and Odontoglossum ringspot virus

2002 ◽  
Vol 147 (10) ◽  
pp. 1943-1954 ◽  
Author(s):  
P. A. Ajjikuttira ◽  
C. L. Lim-Ho ◽  
M. H. Woon ◽  
K. H. Ryu ◽  
C. A. Chang ◽  
...  
Keyword(s):  
Coat Protein ◽  
Genetic Variability ◽  
Mosaic Virus ◽  
Ringspot Virus ◽  
Cymbidium Mosaic Virus ◽  
Odontoglossum Ringspot Virus
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